QbD-Guided RP-HPLC Method Development and Validation for Bisoprolol and Perindopril in Pure and Formulated Dosage Forms

Abstract

A robust and optimized Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method was developed for the simultaneous estimation of Perindopril and Bisoprolol in pharmaceutical dosage forms. Method optimization was performed using a Box-Behnken response surface design, with statistical analysis via ANOVA confirming the significance of resolution and tailing factor models (p < 0.05). Flow rate emerged as a critical factor influencing both responses. Diagnostic plots—including normal residuals, predicted vs. actual values, and 3D surface plots—validated the model's accuracy and predictive capability. System suitability parameters such as resolution (>5), tailing factor (<2), and theoretical plate count (>2000) met all acceptance criteria, ensuring reliable chromatographic performance. The method was validated according to ICH guidelines, demonstrating excellent linearity for Perindopril and Bisoprolol with correlation coefficients (R²) exceeding 0.999. Precision studies showed %RSD values below 2%, confirming repeatability and intermediate precision. Accuracy was supported by recovery rates within 98–102% across multiple concentration levels. Sensitivity was established with low LOD and LOQ values for both analytes, and robustness was confirmed under deliberate variations in chromatographic conditions. Overall, the developed RP-HPLC method is simple, precise, accurate, and robust, making it highly suitable for routine quality control of Perindopril and Bisoprolol in bulk and formulated products.